A DNA Bubble-Mediated Gene Regulation System Based on Thrombin-Bound DNA Aptamers

Jing Wang; Le Yang; Xun Cui; Zhe Zhang; Lichun Dong; Ningzi Guan

doi:10.1021/acssynbio.6b00391

A DNA Bubble-Mediated Gene Regulation System Based on Thrombin-Bound DNA Aptamers

Jing Wang
, Le Yang
, Xun Cui
, Zhe Zhang
, Lichun Dong^*
, Ningzi Guan

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

We describe here a novel approach to enhance the transcription of a target gene in cell-free systems by symmetrically introducing duplex aptamers upstream to a T7 promoter in both the sense and antisense strands of double-stranded plasmids, which leads to the formation of a DNA bubble due to the none-complementary state of the ssDNA region harboring the aptamer sequences. With the presence of thrombins, the DNA bubble would be enlarged due to the binding of aptamers with thrombins. Consequently, the recognition region of the promoter contained in the DNA bubble can be more easily recognized and bound by RNA polymerases, and the separation efficiency of the unwinding region can also be significantly improved, leading to the enhanced expression of the target gene at the transcriptional level. The effectiveness of the proposed gene regulation system was demonstrated by enhancing the expression of gfp and ecaA genes in cell-free systems.

Original language	English
Pages (from-to)	758-765
Number of pages	8
Journal	ACS Synthetic Biology
Volume	6
Issue number	5
DOIs	https://doi.org/10.1021/acssynbio.6b00391
State	Published - 19 May 2017
Externally published	Yes

Keywords

DNA bubble
aptamer
gene expression regulation
promoter

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10.1021/acssynbio.6b00391

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title = "A DNA Bubble-Mediated Gene Regulation System Based on Thrombin-Bound DNA Aptamers",

abstract = "We describe here a novel approach to enhance the transcription of a target gene in cell-free systems by symmetrically introducing duplex aptamers upstream to a T7 promoter in both the sense and antisense strands of double-stranded plasmids, which leads to the formation of a DNA bubble due to the none-complementary state of the ssDNA region harboring the aptamer sequences. With the presence of thrombins, the DNA bubble would be enlarged due to the binding of aptamers with thrombins. Consequently, the recognition region of the promoter contained in the DNA bubble can be more easily recognized and bound by RNA polymerases, and the separation efficiency of the unwinding region can also be significantly improved, leading to the enhanced expression of the target gene at the transcriptional level. The effectiveness of the proposed gene regulation system was demonstrated by enhancing the expression of gfp and ecaA genes in cell-free systems.",

keywords = "DNA bubble, aptamer, gene expression regulation, promoter",

author = "Jing Wang and Le Yang and Xun Cui and Zhe Zhang and Lichun Dong and Ningzi Guan",

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year = "2017",

month = may,

day = "19",

doi = "10.1021/acssynbio.6b00391",

language = "英语",

volume = "6",

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TY - JOUR

T1 - A DNA Bubble-Mediated Gene Regulation System Based on Thrombin-Bound DNA Aptamers

AU - Wang, Jing

AU - Yang, Le

AU - Cui, Xun

AU - Zhang, Zhe

AU - Dong, Lichun

AU - Guan, Ningzi

PY - 2017/5/19

Y1 - 2017/5/19

N2 - We describe here a novel approach to enhance the transcription of a target gene in cell-free systems by symmetrically introducing duplex aptamers upstream to a T7 promoter in both the sense and antisense strands of double-stranded plasmids, which leads to the formation of a DNA bubble due to the none-complementary state of the ssDNA region harboring the aptamer sequences. With the presence of thrombins, the DNA bubble would be enlarged due to the binding of aptamers with thrombins. Consequently, the recognition region of the promoter contained in the DNA bubble can be more easily recognized and bound by RNA polymerases, and the separation efficiency of the unwinding region can also be significantly improved, leading to the enhanced expression of the target gene at the transcriptional level. The effectiveness of the proposed gene regulation system was demonstrated by enhancing the expression of gfp and ecaA genes in cell-free systems.

AB - We describe here a novel approach to enhance the transcription of a target gene in cell-free systems by symmetrically introducing duplex aptamers upstream to a T7 promoter in both the sense and antisense strands of double-stranded plasmids, which leads to the formation of a DNA bubble due to the none-complementary state of the ssDNA region harboring the aptamer sequences. With the presence of thrombins, the DNA bubble would be enlarged due to the binding of aptamers with thrombins. Consequently, the recognition region of the promoter contained in the DNA bubble can be more easily recognized and bound by RNA polymerases, and the separation efficiency of the unwinding region can also be significantly improved, leading to the enhanced expression of the target gene at the transcriptional level. The effectiveness of the proposed gene regulation system was demonstrated by enhancing the expression of gfp and ecaA genes in cell-free systems.

KW - DNA bubble

KW - aptamer

KW - gene expression regulation

KW - promoter

UR - https://www.scopus.com/pages/publications/85019601927

U2 - 10.1021/acssynbio.6b00391

DO - 10.1021/acssynbio.6b00391

M3 - 文章

C2 - 28147483

AN - SCOPUS:85019601927

SN - 2161-5063

VL - 6

SP - 758

EP - 765

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

IS - 5

ER -

A DNA Bubble-Mediated Gene Regulation System Based on Thrombin-Bound DNA Aptamers

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Keywords

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